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Among their likely demands will be the removal of the proposal to create a new EU foreign minister, a controversial idea also included in the original, rejected 2005 EU constitution.

The Labour MPs, led by Ian Davidson, MP for Glasgow South West, may also call on Mr Brown to use a revised treaty to amend the 2004 EU directive on freedom of movement.

The directive is now being blamed for preventing the UK from deporting Learco Chindamo, the man who murdered head teacher Philip Lawrence, back to his native Italy.

Labour went into the 2005 general election promising a referendum on the original EU constitution which was then voted down by Dutch and French voters.

But any attempt to re-open talks now with EU leaders will spark a blazing row, with Brussels insisting that the main issues covered by the treaty agreed at the June EU summit are legally-binding and with only "fine-tuning" now left to be done.

Viscount Etienne Davignon, a former European Commission vice-president, has said that Britain must "stop holding up" other EU members and sign up to the controversial European treaty.

The Prime Minister insists that the new treaty is substantially different from the old constitution and no longer requires a referendum.

Speaking after meeting Angela Merkel, the German Chancellor and main architect of the new treaty, on Wednesday, Mr Brown said: "The proper way to discuss this is in the House of Commons and the House of Lords."

But on Thursday even Gwyneth Dunwoody, the respected and long-serving Labour chairman of the Commons' transport committee, disagreed and called for a referendum in this "rare" case.

The Daily Telegraph's own "let the people decide" petition for a referendum has now been signed by almost 58,000 people - an 8,000 increase in three days.

Labour rebels will be emboldened by the fact that Mr Blair originally refused to promise a referendum on the old constitution plan and was forced to back down.

And Mr Davidson urged the new Prime Minister to abandon Tony Blair's later refusal to hold a vote on the new EU treaty just as he tore up his predecessor's plans for a giant super-casino.

Mr Brown should appeal to other EU leaders to realise that he was now under "domestic pressure" and needed to give way on the referendum, the MP said.

"He could honestly say: a bad boy (Mr Blair) did this and ran away," Mr Davidson told The Daily Telegraph.

Separately, Jim Murphy, the Europe Minister, risked inflaming the rebellion by hitting out at EU "navel-gazing" and denying that the treaty threatened "sweeping new powers" over Britain.

"I am confident that the reform treaty will end the navel-gazing on EU mechanisms and allow the EU to focus on the big issues," he told the ePolitix.com website.

But the concerted action by Eurosceptic Labour MPs came amid mounting pressure from across the political spectrum for a national vote on the EU treaty.

Conservative MPs and Eurosceptic millionaire Paul Sykes joined with Left-wing union leaders to insist that a referendum was the only fair way forward.

Trade union barons made clear they would press ahead with a vote at month's TUC annual congress, demanding that the British people have the final say on the EU treaty.

A motion submitted by the rail workers union RMT and published in the full conference agenda says that the new EU reform Treaty is "substantially the same as the EU Constitution rejected by the French and Dutch electorates in 2005".

It also condemned the blueprint for creating a "centralised government, including an EU president, a Foreign Minister called a High Representative, a diplomatic service and an EU public prosecutor".

But in a sign of conflicting motives even among the unions, Paul Kenny, the GMB general secretary, condemned Britain's opt-outs under the Treaty for failing to give British workers the same rights as "French, German and Spanish workers".

The Open Europe think-tank has sought to embarrass Mr Brown into a U-turn on the referendum by claiming that 170 of Labour's 353 MPs - including Mr Brown himself - were sponsored by or had other links with trade unions now pushing for a referendum.